Cell cDNA synthesis

Material

1. TOPscritTM RT DryMIX (dT18 plus) (enzynomics, Cat#.RT200)
2. TOPscript Reverse Transcriptase
3. TOPscript RT buffer, dNTP mixture
4. RNase inhibitor, Stabilizer
5. Oligo (dT)18
6. DEPC water (Biosesang / cat#. WR2004-050-00)

Method

1. Add DEPC water to total volume 20ul.
2. Add TOPscript RT DryMIX (dT18 plus) 1 tube.
3. Add Template RNA 1uL (100ng).
4. Add Sterile water (RNase free) up to 20uL.
5. Use Total RNA 1ng-5ug; 100ng is used here.
6. For mRNA, use 1ng-250ng.
7. For specific RNA, use 0.01pg-0.5ug.
8. Run cDNA synthesis at 50C for 1hr, then 95C for 5min to inactivate the reaction.
9. Store at 20C.